Psychostimulant is a type of psychoactive substances that stimulate central and peripheral nervous system as well as induce drug dependence.A series of studies indicate that monoamine especially dopamine plays an important role in behavior and drug dependence,moreover,dopamine transporter(DAT)controls homeostasis of dopamine in neuron and transmission of dopamine pathways.Thus DAT might play an important role in the reward and behavioral stimulation of psychostimulant.

Objective To construct NT4-GFP-Ant fusional reporting gene and the vector of NT4-GFP-Ant recombinant adeno-associated virus(AAV).Methods The GFP gene was cloned by using PCR and T-vector cloning method.The positive clone was identified by the restriction enzymes,and then the cloned amplified fragments were sequenced and analyzed.The resulting gene of GFP and Ant,PBV220/NT4 were connected by DNA ligase,and thus PBV220/NT4-GFP-Ant was constructed,then the NT4-GFP-Ant fragment was gained and identified by the restriction enzymes.The resulting gene of NT4-GFP-Ant fragment was inserted into the EcoRⅠ-BamHⅠsite of vector plasmid pSSHG to construct the vector of NT4-GFP-Ant recombinant AAV.Results A 730 bp fragment of DNA was gained when T-easy/GFP was cut by the restriction enzyme EcoRⅠ.The cloned GFP gene was coincident with the sequence in GenBank.A 1 000 bp fragment of DNA was gained when pBV220/NT4-GFP-Ant was cut by the restriction enzymes BamHⅠ/EcoRⅠ.A 1 000 bp fragment of DNA was gained when PSSHG/NT4-GFP-Ant was cut by the restriction enzymes BamHⅠ/EcoRⅠ.Conclusion GFP gene is cloned successfully,NT4-GFP-Ant gene and PSSHG/NT4-GFP-Ant recombinant AAV vector are constructed successfully.

AIM: To investigate the variation and distribution of abnormaly methylated p15 INK4B in myelodysplastic syndrome (MDS) and its subgroups. METHODS: The abnormal methylation of p15 INK4B in 32 cases with MDS was studied using methylation-specific PCR (MSP). RESULTS: The positive rate of abnormal methylation of p15 INK4B was about 50% in MDS. The ratios in subtypes were: 0% (0/6) in RA,20% (1/5) in RA-S,57.1% (4/7) in RAEB,74.1% (5/7) in CMML,85.7% (6/7) in RAEB-t, respectively.It was worth noticing that 4 cases represented abnormal methylation of p15INK4B during their transformation and progression into AML. CONCLUSION: The abnormal methylation in p15 INK4B might be one of the causes of MDS, which was related to pathologial process of MDS.Every subtype was not solitary classification completely. Abnormal methylation of p15 INK4B was apt to occur accompanying the progression and transformation of the subtypes.

To study the action, characteristics and expression of high methylation of p15(INK4B) and p16(INK4A) genes in multiple myeloma (MM), the sensitive methylation specific PCR method was employed to detect the hypermethylation of p15(INK4B) and p16(INK4A) in 24 patients with MM. Results showed that the methylation incidence of p15(INK4B) and p16(INK4A) genes were 70.8% (17/24) and 58.3% (14/24) in the MM patients, with the products of 148 bp and 150 bp fragments, respectively. The methylation of p15(INK4B) and p16(INK4A) genes were simultaneously happened in MM patients of plasmocytoma type with two cases at II phase and two cases at III phase. The simultaneous non-methylation of p15(INK4B) and p16(INK4A) genes were founded in five cases of MM patients, all of the tumor cells were of small plasmocyte type with mature differention. Conclusion suggested that there were high incidence of methylation of p15(INK4B) and p16(INK4A) genes in patients with MM. Hypermethylation can be detected in the early stage of disease, which was associated with its progress. It indicated a bad prognosis when methylation happended simultaneously in the two genes. Methylation of p15(INK4B) and p16(INK4A) genes may be related to the pathogeny of MM.